Crystal forms of azithromycin

ABSTRACT

The invention relates to novel crystal forms of azithromycin, an antibiotic useful in the treatment of infections.

BACKGROUND OF THE INVENTION

This invention relates to crystal forms of azithromycin. Azithromycin issold commercially and is an effective antibiotic in the treatment of abroad range of bacterial infections. The crystal forms of this inventionare likewise useful as antibiotic agents in mammals, including man, aswell as in fish and birds.

Azithromycin has the following structural formula:

Azithromycin is described and claimed in U.S. Pat. Nos. 4,517,359 and4,474,768. It is also known as9-deoxo-9a-aza-9a-methyl-9a-homoerythomycin A.

Other patents or patent applications which directly or indirectly coverazithromycin include: EP 298,650 which claims azithromycin dihydrate;U.S. Pat. No. 4,963,531 which claims a method of treating a strain ofToxoplasma gondi species; U.S. Pat. No. 5,633,006 which claims achewable tablet or liquid suspension pharmaceutical composition havingreduced bitterness; U.S. Pat. No. 5,686,587 which claims an intermediateuseful in the preparation of azithromycin; U.S. Pat. No. 5,605,889 whichclaims an oral dosage form that reduces the “food effect” associatedwith the administration of azithromycin; U.S. Pat. No. 6,068,859 whichclaims a controlled dosage form containing azithromycin; U.S. Pat. No.5,498,699 which claims a composition containing azithromycin incombination with bivalent or trivalent metals; EP 925,789 which claims amethod of treating eye infections; Chinese patent application CN1123279A which relates to water soluble salts of azithromycin; Chinesepatent application CN 1046945C which relates to azithromycin sodiumdihydrogenphosphate double salts; Chinese patent application CN 1114960Awhich relates to azithromycin crystals, Chinese patent application CN1161971A which relates to azithromycin crystals; Chinese patentapplication CN 1205338A which relates to a method of preparing watersoluble salts of azithromycin; International Publication WO 00/32203which relates to an ethanolate of azithromycin; and European patentapplication EP 984,020 which relates to an azithromycin monohydrateisopropanol clathrate.

SUMMARY OF THE INVENTION

The present invention relates to crystal forms of azithromycin. As usedherein, the term “crystal form(s)” or “form(s)”, unless otherwise noted,means one or more crystal forms of azithromycin.

In particular, the present invention relates to a crystal form ofazithromycin wherein said crystal form is selected from forms C, D, E,F, G, H, J, M, N, O, P, Q and R wherein said forms are as definedherein. Forms F, G, H, J, M, N, O, and P belong to family I azithromycinand belong to a monoclinic P2₁ space group with cell dimensions ofa=16.3±0.3 Å, b=16.2±0.3 Å, c=18.4±0.3 Å and beta=109±2°. Forms C, D, Eand R belong to family II azithromycin and belong to an orthorhombicP2₁2₁2₁ space group with cell dimensions of a=8.9±0.4 Å, b=12.3±0.5 Åand c=45.8±0.5 Å. Form Q is distinct from families I and II.

Form F azithromycin is of the formula C₃₈H₇₂N₂O₁₂.H₂O.0.5C₂H₅OH in thesingle crystal structure, being azithromycin monohydrate hemi-ethanolsolvate. Form F is further characterized as containing 2-5% water and1-4% ethanol by weight in powder samples and having powder X-raydiffraction 2θ peaks as defined in Table 9. The ¹³C ssNMR (solid stateNuclear Magnetic Resonance) spectrum of form F has two chemical shiftpeaks at approximately 179±1 ppm, those being 179.5±0.2 ppm and178.6±0.2 ppm, a set of five peaks between 6.4 to 11.0 ppm, and ethanolpeaks at 58.0±0.5 ppm and 17.2±0.5 ppm. The solvent peaks can be broadand relatively weak in intensity.

The invention also relates to substantially pure form F azithromycin,form F azithromycin substantially free of form G azithromycin and form Fazithromycin substantially free of azithromycin dihydrate.

The invention further relates to methods of preparing form Fazithromycin by treating azithromycin with ethanol to completedissolution at 40-70° C. and cooling with reduction of ethanol oraddition of water to effect crystallization. Also included are methodsof making substantially pure form F azithromycin, form F azithromycinsubstantially free of form G azithromycin and form F azithromycinsubstantially free of azithromycin dihydrate.

Form G azithromycin is of the formula C₃₈H₇₂N₂O₁₂.1.5H₂O in the singlecrystal structure, being azithromycin sesquihydrate. Form G is furthercharacterized as containing 2.5-6% water and <1% organic solvent(s) byweight in powder samples and having powder X-ray diffraction 2θ peaks asdefined in Table 9. The ¹³C ssNMR spectrum of form G has one chemicalshift peak at approximately 179±1 ppm, being a peak at 179.5±0.2 ppm(splitting <0.3 ppm may present), and a set of five peaks between 6.3 to11.0 ppm.

The invention also relates to substantially pure form G azithromycin,and form G azithromycin substantially free of azithromycin dihydrate.

The invention further relates to methods of preparing substantially pureform G azithromycin, and form G azithromycin substantially free ofazithromycin dihydrate by treating azithromycin with a mixture ofmethanol and water or acetone and water to complete dissolution at40-60° C. and cooling to effect crystallization.

Form H azithromycin is of the formula C₃₈H₇₂N₂O₁₂.H₂O.C₃H₈O₂ beingazithromycin monohydrate hemi-1,2 propanediol solvate.

Form J azithromycin is of the formula C₃₈H₇₂N₂O₁₂.H₂O.0.5C₃H₇OH in thesingle crystal structure, being azithromycin monohydrate hemi-n-propanolsolvate. Form J is further characterized as containing 2-5% water and1-5% 1-propanol by weight in powder samples and having powder X-raydiffraction 20 peaks as defined in Table 9. The ¹³C ssNMR spectrum ofform J has two chemical shift peaks at approximately 179±1 ppm, thosebeing 179.6±0.2 ppm and 178.4±0.2 ppm, a set of five peaks between 6.6to 11.7 ppm and an n-propanol peak at 25.2±0.4 ppm. The solvent peak canbe broad and relatively weak in intensity.

The invention further relates to methods of preparing form J by treatingazithromycin with n-propanol to complete dissolution at 25-55° C. andcooling with addition of water to effect crystallization.

Form M azithromycin is of the formula C₃₈H₇₂N₂O₁₂.H₂O.0.5C₃H₇OH, beingazithromycin monohydrate hemi-isopropanol solvate. Form M is furthercharacterized as containing 2-5% water and 1-4% 2-propanol by weight inpowder samples and having powder X-ray diffraction 2θ peaks as definedin Table 9. The ¹³C ssNMR spectrum of form M has one chemical shift peakat approximately 179±1 ppm, being 179.6±0.2 ppm, a peak at 41.9±0.2 ppmand a set of six peaks between 6.9 to 16.4 ppm and an isopropanol peakat 26.0±0.4 ppm. The solvent peak can be broad and relatively weak inintensity.

The invention also relates to substantially pure form M azithromycin,form M azithromycin substantially free of form G azithromycin and form Mazithromycin substantially free of azithromycin dihydrate.

The invention further relates to methods of preparing substantially pureform M azithromycin, form M azithromycin substantially free of form Gazithromycin and form M azithromycin substantially free of azithromycindihydrate by treating azithromycin with isopropanol to completedissolution at 40-60° C. and reduction of isopropanol followed bycooling or cooling followed by addition of water to effectcrystallization.

Form N azithromycin is a mixture of isomorphs of Family I. The mixturemay contain variable percentages of isomorphs, F, G, H, J, M and others,and variable amounts of water and organic solvents, such as ethanol,isopropanol, n-propanol, propylene glycol, acetone, acetonitrile,butanol, pentanol, etc. The weight percent of water can range from 1-5%and the total weight percent of organic solvents can be 2-5% with eachsolvent content of 0.5 to 4%. The samples of form N display allcharacteristic peaks of members of Family I in various proportions. FormN may be characterized as ‘mixed crystals’ or “crystalline solidsolutions' of Family I isomorphs.

Form N displays chemical shifts as a combination of isomorphs in FamilyI. The peaks may vary in chemical shift ppm within ±0.2 ppm and inrelative intensities and width due to the mixing of variable proportionof isomorphs contained in the form N crystalline solid solution.

Form P azithromycin is of the formula C₃₈H₇₂N₂O₁₂.H₂O.0.5C₅H₁₂O beingazithromycin monohydrate hemi-n-pentanol solvate.

Form Q azithromycin is of the formula C₃₈H₇₂N₂O₁₂.H₂O.0.5C₄H₈O beingazithromycin monohydrate hemi-tetrahydrofuran solvate.

Form R azithromycin is of the formula C₃₈H₇₂N₂O₁₂.H₂O.C₅H₁₂O beingazithromycin monohydrate mono-methyl tert-butyl ether solvate.

Form D azithromycin is of the formula C₃₈H₇₂N₂O₁₂.H₂O.C₆H₁₂ in itssingle crystal structure, being azithromycin monohydrate monocyclohexanesolvate. Form D is further characterized as containing 2-6% water and3-12% cyclohexane by weight in powder samples and having representativepowder X-ray diffraction 2θ peaks as defined in Table 9. The ¹³C ssNMRspectrum of form D displays has one chemical shift peak at approximately179±1 ppm, being 178.1±0.2 ppm and peaks at 103.9±0.2 ppm, 95.1±0.2 ppm,84.2±0.2 ppm, and a set of 3 peaks between 8.4 to 11 ppm.

The invention further relates to methods of preparing form D byslurrying azithromycin dihydrate with cyclohexane.

Form E azithromycin is of the formula C₃₈H₇₂N₂O₁₂.H₂O.C₄H₈O beingazithromycin monohydrate mono-tetrahydrofuran solvate.

The invention further relates to azithromycin in an amorphous state anda method of preparing amorphous azithromycin that comprises the removalof water and/or solvents from the azithromycin crystal lattice. TheX-ray diffraction powder pattern for amorphous azithromycin displays nosharp 2θ peaks but has two broad rounded peaks. The first peak occursbetween 40 and 130. The second peak occurs between 13° and 250.

The invention also relates to pharmaceutical compositions for thetreatment of a bacterial infection or a protozoa infection in a mammal,fish, or bird which comprises a therapeutically effective amount of thecrystalline compounds referred to above, or amorphous azithromycin, anda pharmaceutically acceptable carrier.

The invention also relates to a method of treating a bacterial infectionor a protozoa infection in a mammal, fish, or bird which comprisesadministering to said mammal, fish or bird a therapeutically effectiveamount of the crystalline compounds referred to above, or amorphousazithromycin.

The present invention also relates to methods of preparing crystal formsof azithromycin which comprise the slurrying of azithromycin in anappropriate solvent or the dissolution of azithromycin in a heatedorganic solvent or organic solvent/water solution and precipitating thecrystalline azithromycin by cooling the solution with reduction ofsolvent volume or by dissolving azithromycin in a solvent or solventmixture and precipitating crystalline azithromycin by the addition ofwater to the solution. Azithromycin in amorphous state is prepared byheating crystalline azithromycin in a vacuum.

The term “treatment”, as used herein, unless otherwise indicated, meansthe treatment or prevention of a bacterial infection or protozoainfection as provided in the method of the present invention, includingcuring, reducing the symptoms of or slowing the progress of saidinfection. The terms “treat” and “treating” are defined in accord theforegoing term “treatment”.

The term “substantially free” when referring to a designated crystallineform of azithromycin means that there is less than 20% (by weight) ofthe designated crystalline form(s) present, more preferably, there isless than 10% (by weight) of the designated form(s) present, morepreferably, there is less than 5% (by weight) of the designated form(s)present, and most preferably, there is less than 1% (by weight) of thedesignated crystalline form(s) present. For instance, form Fazithromycin substantially free of azithromycin dihydrate means form Fwith 20% (by weight) or less of azithromycin dihydrate, more preferably,10% (by weight) or less of azithromycin dihydrate, most preferably, 1%(by weight) of azithromycin dihydrate.

The term “substantially pure” when referring to a designated crystallineform of azithromycin means that the designated crystalline form containsless than 20% (by weight) of residual components such as alternatepolymorphic or isomorphic crystalline form(s) of azithromycin. It ispreferred that a substantially pure form of azithromycin contain lessthan 10% (by weight) of alternate polymorphic or isomorphic crystallineforms of azithromycin, more preferred is less than 5% (by weight) ofalternate polymorphic or isomorphic crystalline forms of azithromycin,and most preferably less than 1% (by weight) of alternate polymorphic orisomorphic crystalline forms of azithromycin.

The term “substantially in the absence of azithromycin dihydrate” whenreferring to bulk crystalline azithromycin or a composition containingcrystalline azithromycin means the crystalline azithromycin containsless than about 5% (by weight) azithromycin dihydrate, more preferablyless than about 3% (by weight) azithromycin dihydrate, and mostpreferably less than 1% (by weight) azithromycin dihydrate.

As used herein, unless otherwise indicated, the term “bacterialinfection(s)” or “protozoa infection” includes bacterial infections andprotozoa infections and diseases caused by such infections that occur inmammals, fish and birds as well as disorders related to bacterialinfections and protozoa infections that may be treated or prevented byadministering antibiotics such as the compound of the present invention.Such bacterial infections and protozoa infections and disorders relatedto such infections include, but are not limited to, the following:pneumonia, otitis media, sinusitus, bronchitis, tonsillitis, andmastoiditis related to infection by Streptococcus pneumoniae,Haemophilus influenzae, Moraxella catarrhalis, Staphylococcus aureus, orPeptostreptococcus spp.; pharynigitis, rheumatic fever, andglomerulonephritis related to infection by Streptococcus pyogenes,Groups C and G streptococci, Clostridium diptheriae, or Actinobacillushaemolyticum; respiratory tract infections related to infection byMycoplasma pneumoniae, Legionella pneumophila, Streptococcus pneumoniae,Haemophilus influenzae, or Chlamydia pneumoniae; uncomplicated skin andsoft tissue infections, abscesses and osteomyelitis, and puerperal feverrelated to infection by Staphylococcus aureus, coagulase-positivestaphylococci (i.e., S. epidermidis, S. hemolyticus, etc.),Streptococcus pyogenes, Streptococcus agalactiae, Streptococcal groupsC-F (minute-colony streptococci), viridans streptococci, Corynebacteriumminutissimum, Clostridium spp., or Bartonella henselae; uncomplicatedacute urinary tract infections related to infection by Staphylococcussaprophyticus or Enterococcus spp.; urethritis and cervicitis; andsexually transmitted diseases related to infection by Chlamydiatrachomatis, Haemophilus ducreyi, Treponema pallidum, Ureaplasmaurealyticum, or Neiserria gonorrheae; toxin diseases related toinfection by S. aureus (food poisoning and Toxic shock syndrome), orGroups A, B, and C streptococci; ulcers related to infection byHelicobacter pylori; systemic febrile syndromes related to infection byBorrelia recurrentis; Lyme disease related to infection by Borreliaburgdorferi, conjunctivitis, keratitis, and dacrocystitis related toinfection by Chlamydia trachomatis, Neisseria gonorrhoeae, S. aureus, S.pneumoniae, S. pyogenes, H. influenzae, or Listeria spp.; disseminatedMycobacterium avium complex (MAC) disease related to infection byMycobacterium avium, or Mycobacterium intracellulare; gastroenteritisrelated to infection by Campylobacter jejuni; intestinal protozoarelated to infection by Cryptosporidium spp.; odontogenic infectionrelated to infection by viridans streptococci; persistent cough relatedto infection by Bordetella pertussis; gas gangrene related to infectionby Clostridium perfringens or Bacteroides spp.; and atherosclerosisrelated to infection by Helicobacter pylori or Chlamydia pneumoniae.Also included are atherosclerosis and malaria. Bacterial infections andprotozoa infections and disorders related to such infections that may betreated or prevented in animals include, but are not limited to, thefollowing: bovine respiratory disease related to infection by P. haem.,P. multocida, Mycoplasma bovis, or Bordetella spp.; cow enteric diseaserelated to infection by E. coli or protozoa (i.e., coccidia,cryptosporidia, etc.); dairy cow mastitis related to infection by Staph.aureus, Strep. uberis, Strep. agalactiae, Strep. dysgalactiae,Klebsiella spp., Corynebacterium, or Enterococcus spp.; swinerespiratory disease related to infection by A. pleuro., P. multocida, orMycoplasma spp.; swine enteric disease related to infection by E. coli,Lawsonia intracellularis, Salmonella, or Serpulina hyodyisinteriae; cowfootrot related to infection by Fusobacterium spp.; cow metritis relatedto infection by E. coli; cow hairy warts related to infection byFusobacterium necrophorum or Bacteroides nodosus; cow pink-eye relatedto infection by Moraxella bovis; cow premature abortion related toinfection by protozoa (i.e. neosporium); urinary tract infection in dogsand cats related to infection by E. coli; skin and soft tissueinfections in dogs and cats related to infection by Staph. epidermidis,Staph. intermedius, coagulase neg. Staph. or P. multocida; and dental ormouth infections in dogs and cats related to infection by Alcaligenesspp., Bacteroides spp., Clostridium spp., Enterobacter spp.,Eubacterium, Peptostreptococcus, Porphyromonas, or Prevotella. Otherbacterial infections and protozoa infections and disorders related tosuch infections that may be treated or prevented in accord with themethod of the present invention are referred to in J. P. Sanford et al.,“The Sanford Guide To Antimicrobial Therapy,” 26th Edition,(Antimicrobial Therapy, Inc., 1996).

The present invention also includes isotopically-labeled compoundswherein one or more atoms are replaced by an atom having an atomic massor mass number different from the atomic mass or mass number usuallyfound in nature. Examples of isotopes that can be incorporated intocompounds of the invention include isotopes of hydrogen, carbon,nitrogen, oxygen, phosphorous, sulfur, fluorine and chlorine, such as²H, ³H, ¹³C, ¹⁴C, ¹⁵N, ¹⁸O, and ¹⁷O. Such radiolabelled andstable-isotopically labelled compounds are useful as research ordiagnostic tools.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a calculated powder X-ray diffraction pattern of azithromycinform A. The scale of the abscissa is degrees 2-theta (2 θ). The ordinateis the intensity in counts.

FIG. 2 is an experimental powder X-ray diffraction pattern ofazithromycin form A. The scale of the abscissa is in degrees 2-theta (2θ). The ordinate is the intensity in counts.

FIG. 3 is an overlay of FIGS. 1 and 2 with the calculated diffractionpatterns of azithromycin form A (FIG. 1) on the bottom and theexperimental diffraction pattern of azithromycin form A (FIG. 2) on thetop. The scale of the abscissa is in degrees 2-theta (2 θ). The ordinateis the intensity in counts.

FIG. 4 is a calculated powder X-ray diffraction pattern of azithromycinform C. The scale of the abscissa is in degrees 2-theta (2 θ). Theordinate is the intensity in counts.

FIG. 5 is a calculated powder X-ray diffraction pattern of azithromycinform D. The scale of the abscissa is in degrees 2-theta (2 θ). Theordinate is the intensity in counts.

FIG. 6 is an experimental powder X-ray diffraction pattern ofazithromycin form D. The scale of the abscissa is in degrees 2-theta (2θ). The ordinate is the intensity in counts.

FIG. 7 is an overlay of FIGS. 5 and 6 with the calculated diffractionpattern of azithromycin form D (FIG. 5) on the bottom and theexperimental diffraction pattern of azithromycin form D (FIG. 6) on thetop. The scale of the abscissa is in degrees 2-theta (2 θ). The ordinateis the intensity in counts.

FIG. 8 is a calculated powder X-ray diffraction pattern of azithromycinform E. The scale of the abscissa is in degrees 2-theta (2 θ). Theordinate is the intensity in counts.

FIG. 9 is a calculated powder X-ray diffraction pattern of azithromycinform F. The scale of the abscissa is in degrees 2-theta (2 θ). Theordinate is the intensity in counts.

FIG. 10 is an experimental powder X-ray diffraction pattern ofazithromycin form F. The scale of the abscissa is in degrees 2-theta (2θ). The ordinate is the intensity in counts.

FIG. 11 is an overlay of FIGS. 9 and 10 with the calculated diffractionpattern of azithromycin form F (FIG. 9) on the bottom and theexperimental diffraction pattern of azithromycin form F (FIG. 10) on thetop. The scale of the abscissa is in degrees 2-theta (2 θ). The ordinateis the intensity in counts.

FIG. 12 is a calculated powder X-ray diffraction pattern of azithromycinform G. The scale of the abscissa is in degrees 2-theta (2 θ). Theordinate is the intensity is counts.

FIG. 13 is an experimental powder X-ray diffraction pattern ofazithromycin form G. The scale of the abscissa is in degrees 2-theta (2θ). The ordinate is the intensity in counts.

FIG. 14 is an overlay of FIGS. 12 and 13 with the calculated diffractionpattern of azithromycin form G (FIG. 12) on the bottom and theexperimental diffraction pattern of azithromycin form G (FIG. 13) on thetop. The scale of the abscissa is in degrees 2-theta (2 θ). The ordinateis the intensity in counts.

FIG. 15 is a calculated powder X-ray diffraction pattern of azithromycinform J. The scale of the abscissa is in degrees 2-theta (2 θ). Theordinate is the intensity in counts.

FIG. 16 is an experimental powder X-ray diffraction pattern ofazithromycin form J. The scale of the abscissa is in degrees 2-theta (2θ). The ordinate is the intensity in counts.

FIG. 17 is an overlay of FIGS. 15 and 16 with the calculated diffractionpattern of azithromycin form J (FIG. 15) on the bottom and theexperimental diffraction pattern of azithromycin form J (FIG. 16) on thetop. The scale of the abscissa is in degrees 2-theta (2 θ). The ordinateis the intensity in counts.

FIG. 18 is an experimental powder X-ray diffraction pattern ofazithromycin form M. The scale of the abscissa is in degrees 2-theta (2θ). The ordinate is the intensity in counts.

FIG. 19 is an experimental powder X-ray diffraction pattern ofazithromycin form N. The scale of the abscissa is in degrees 2-theta (2θ). The ordinate is the intensity in counts.

FIG. 20 is an experimental powder X-ray diffraction pattern of amorphousazithromycin. The scale of the abscissa is in degrees 2-theta (2 θ). Theordinate is the intensity in counts.

FIG. 21 is a ¹³C solid state NMR spectrum of azithromycin form A.

FIG. 22 is a ¹³C solid state NMR spectrum of azithromycin form D.

FIG. 23 is a ¹³C solid state NMR spectrum of azithromycin form F.

FIG. 24 is a ¹³C solid state NMR spectrum of azithromycin form G.

FIG. 25 is a ¹³C solid state NMR spectrum of azithromycin form J.

FIG. 26 is a ¹³C solid state NMR spectrum of azithromycin form M.

FIG. 27 is a ¹³C solid state NMR spectrum of azithromycin form N.

FIG. 28 is a ¹³C solid state NMR spectrum of amorphous azithromycin.

FIG. 29 is a ¹³C solid state NMR spectrum of a pharmaceutical tabletcontaining form G azithromycin.

FIG. 30 is an experimental powder X-ray diffraction pattern ofazithromycin form Q. The scale of the abscissa is in degrees 2-theta (2θ). The ordinate is the intensity in counts.

FIG. 31 is an experimental powder X-ray diffraction pattern ofazithromycin form R. The scale of the abscissa is in degrees 2-theta (2θ). The ordinate is the intensity in counts.

FIG. 32 is a ¹³C solid state NMR spectrum of azithromycin form H.

FIG. 33 is a ¹³C solid state NMR spectrum of azithromycin form R.

DETAILED DESCRIPTION OF THE INVENTION

Azithromycin has been found to exist in different crystalline forms. Adihydrate, form A, and a non-stroichiometric hydrate, form B, aredisclosed in European Patent EP 298 650 and U.S. Pat. No. 4,512,359,respectively. Sixteen other forms have been discovered, namely forms C,D, E, F, G, H, I, J, K, L, M, N, O, P, Q and R. These forms are eitherhydrates or hydrate/solvates of azithromycin free base. Forms L and Kare the metastable lower hydrate forms of A, detected at hightemperature. Crystal structures of forms A, C, D, E, F, G, H, J and Ohave been solved. The structural data of these crystal forms are givenbelow: TABLE 1 Crystallographic data of azithromycin form A. Form AEmpirical formula C₃₈H₇₂N₂O₁₂.2H₂O Formula weight 785.2 Crystal size(mm) 0.19 × 0.24 × 0.36 Space group P2₁2₁2₁ orthorhombic Unit celldimensions a = 14.735 (5) Å b = 16.844 (7) Å c = 17.81 (1) Å α = 90° β =90° γ = 90° Z (per formula)  4 Density (g/cm³)  1.18 R  0.060

TABLE 2 Crystallographic data of azithromycin form C. Form C Empiricalformula C₃₈H₇₂N₂O₁₂.H₂O Formula weight 767.15 Crystal size (mm) 0.16 ×0.16 × 0.19 Space group P2₁2₁2₁ orthorhombic Unit cell dimensions a =8.809 (3) Å b = 12.4750 (8) Å c = 45.59 (3) Å α = 90° β = 90° γ = 90° Z(per formula)  4 Density (g/cm³)  1.01 R  0.106

TABLE 3 Crystallographic data of azithromycin form D. Form D Empiricalformula C₃₈H₇₂N₂O₁₂.H₂O.C₆H₁₂ Formula weight 851.15 Crystal size (mm)0.52 × 0.32 × 0.16 Space group P2₁2₁2₁orthorhombic Unit cell dimensionsa = 8.8710 (10) Å b = 12.506 (2) Å c = 45.697 (7) Å α = 90° β = 90° γ =90° Z (per formula)  4 Density (g/cm³)  1.12 R  0.0663

TABLE 4 Crystallographic data of azithromycin form E. Form E Empiricalformula C₃₈H₇₂N₂O₁₂.H₂O.C₄H₈O Formula weight 839.2 Crystal size (mm)0.17 × 0.19 × 0.20 Space group P2₁2₁2₁ orthorhombic Unit cell dimensionsa = 8.869 (3) Å b = 12.086 (3) Å c = 46.00 (1) Å α = 90° β = 90° γ = 90°Z (per formula)  4 Density (g/cm³)  1.13 R  0.087

TABLE 5 Crystallographic data of azithromycin form F. Form F Empiricalformula C₃₈H₇₂N₂O₁₂.H₂O.0.5C₂H₆O Crystal size (mm) 0.14 × 0.20 × 0.24Formula weight 790.2 Space group P2₁ monoclinic Unit cell dimensions a =16.281 (2) Å b = 16.293(1) Å c = 18.490 (3) Å α = 90° β = 109.33(1)° γ =90° Z (per formula)  4 Density (g/cm³)  1.13 R  0.0688

TABLE 6 Crystallographic data of azithromycin form G. Form G FormulaC₃₈H₇₂N₂O₁₂.1.5H₂O Formula weight 776.0 Crystal size (mm) 0.04 × 0.20 ×0.24 Space group P2₁ monoclinic Unit cell dimensions a = 16.4069(8) Å b= 16.2922(8) Å c = 18.3830 (9) Å α = 90° β = 110.212(2)° γ = 90° Z (performula)  4 Density (g/cm³)  1.12 R  0.0785

TABLE 7 Crystallographic data of azithromycin form H. Form H Empiricalformula C₃₈H₇₂N₂O₁₂.H₂O.0.5C₃H₈O₂ Crystal size (mm) 0.14 × 0.20 × 0.24Formula weight 805.0 Space group P2₁ monoclinic Unit cell dimensions a =16.177 (1) Å b = 16.241 (2) Å c = 18.614 (1) Å α = 90° β = 108.34 (1)° γ= 90° Z (per formula)  4 Density (g/cm³)  1.15 R  0.0687

TABLE 8 Crystallographic data of azithromycin form J. Form J FormulaC₃₈H₇₂N₂O₁₂.H₂O.0.5C₃H₈O Formula weight 796.0 Crystal size (mm) 0.40 ×0.36 × 0.20 Space group P2₁ monoclinic Unit cell dimensions a =16.191(6) Å b = 16.237(10) Å c = 18.595(14) Å α = 90° β = 108.92(4)° γ =90° Z (per formula)  4 Density (g/cm³)  1.14 R  0.0789

TABLE 8A Crystallographic data of azithromycin form O. Form O FormulaC₃₈H₇₂N₂O₁₂.0.5H₂O.0.5C₄H₁₀O Formula weight 795.04 Crystal size (mm)0.40 × 0.36 × 0.20 Space group P2₁ monoclinic Unit cell dimensions a =16.3602(11) Å b = 16.2042(11) Å c = 18.5459(12) Å α = 90° β =109.66(10)° γ = 90° Z (per formula)  4 Density (g/cm³)  1.14 R  0.0421

Among these sixteen crystal forms, two isomorphic families areidentified. Family I includes forms F, G, H, J, M, N, O, and P. FamilyII includes forms C, D, E and R. Form Q is distinct from families I andII. The forms within a family are isomorphs that crystallize in the samespace group with slight variation of cell parameters and comprisechemically related structures but different elemental composition. Inthis case, the variation in chemical composition among the isomorphsarises from incorporation of different water/solvent molecules.Consequently, the isomorphs display similar but non-identical X-raydiffraction patterns and solid-state NMR spectra (ssNMR). Othertechniques such as near infrared spectroscopy (NIR), differentialscanning calorimetry (DSC), gas chromatography (GC), thermalgravimetricanalysis (TGA), or thermalgravimetric analysis/infrared spectroscopyanalysis (TG-IR), Karl Fischer water analysis (KF) and molecularmodeling/visualization provide data for affirmative identification ofisomorphs. Dehydration/desolvation temperatures were determined by DSCwith a heating rate of 5° C./min

Form C: This crystal form was identified from a single crystal structure(Table 2)—a monohydrate of azithromycin. It has the space group ofP2₁2₁2₁ and similar cell parameters as that of forms D and E; therefore,it belongs to Family II isomorphs. Its calculated powder pattern issimilar to that of forms D and E.

Form D: Form D was crystallized from cyclohexane. The single crystalstructure of form D shows a stoichiometry of amonohydrate/monocyclohexane solvate of azithromycin (Table 3).Cyclohexane molecules were found to be disordered in the crystallattice. From single crystal data, the calculated water and cyclohexanecontent of form D is 2.1 and 9.9%, respectively. Both the powder patternand the calculated powder pattern of form D are similar to those offorms C and E. The powder samples of form D showed adesolvation/dehydration endotherm with an onset temperature of about 87°C. and a broad endotherm between 200-280° C. (decomposition) in DSCanalysis at 5° C./min from 30-300° C.

Form D is prepared by slurrying azithromycin in cyclohexane for 2-4days. The solid form D azithromycin is collected by filtration anddried.

Form E: Form E was obtained as a single crystal collected in a THF/watermedium. It is a monohydrate and mono-THF solvate by single crystalanalysis (Table 4). By its single crystal structure, the calculated PXRDpattern is similar to that of form C and form D making it a family IIisomorph.

Form E is prepared by dissolving azithromycin in THF (tetrahydrofuran).Diffusing water vapor through saturated azithromycin THF solution overtime yields crystals of Form E.

Form F: The single crystal of form F crystallized in a monoclinic spacegroup, P2₁, with the asymmetric unit containing two azithromycin, twowaters, and one ethanol, as a monohydrate/hemi-ethanolate (Table 5). Itis isomorphic to all family I azithromycin crystalline forms. Thecalculated PXRD pattern of this form is similar to those of other familyI isomorphs. The theoretical water and ethanol contents are 2.3 and2.9%, respectively. The powder samples show a dehydration/desolvationendotherm at an onset temperature between 110-125° C. Form F is preparedby dissolving azithromycin in ethanol (1-3 volumes by weight) at atemperature of about 50-70° C. Upon complete dissolution, the solutionis cooled to subambient temperature to cause precipitation. The volumeof ethanol can be reduced by vacuum distillation with stirring for 1-2hours to increase the yield. Alternatively, water (optionally chilled to0-20° C.) about 0.1-2 volume can be added with collection of solidswithin 30 minute after water addition. Cooling the ethanol solution ofazithromycin prior to the addition of water to below below 20° C.,preferably below 15° C., more preferably below 10, and most preferably5° C. results in substantially pure azithromycin form F. The solid formF azithromycin is collected by filtration and dried.

Form G: The single crystal structure of form G consists of twoazithromycin molecules and three water molecules per asymmetric unit(Table 6). This corresponds to a sesquihydrate with a theoretical watercontent of 3.5%. The water content of powder samples of form G rangesfrom about 2.5 to about 6%. The total residual organic solvent is lessthan 1% of the corresponding solvent used for crystallization, which iswell below stoichiometric quantities of solvate. This form dehydrateswith an onset temperature of about 110-120° C.

Form G may be prepared by adding azithromycin to a premixed organicsolvent/water mixture (1/1 by volume), where the organic solvent can bemethanol, acetone, acetonitrile, ethanol or isopropanol. The mixture isstirred and heated to an elevated temperature, e.g. 45-55° C. for 4-6hours to cause dissolution. Precipitation occurs during cooling toambient temperature. The solid form G azithromycin is collected byfiltration and dried.

Form H: This crystal form is a monohydrate/hemi-propylene glycol solvateof azithromycin free base (Table 7). It was isolated from a formulationsolution containing propylene glycol. The crystal structure of form H isisomorphic to crystal forms of Family I.

Azithromycin form H is prepared by dissolving azithromycin dihydrate in6 volumes of propylene glycol. To the resulting propylene glycolsolution of azithromycin, 2 volumes of water is added and precipitationoccurrs. The slurry is stirred for 24 hours and the solids are filteredand air-dried at ambient temperature to afford crystalline Form H.

Form J: Form J is a monohydrate/hemi n-propanol solvate (Table 8). Thecalculated solvent content is about 3.8% n-propanol and about 2.3%water. The experimental data shows from about 2.5 to about 4.0%n-propanol and from about 2.5 to about 3% water content for powdersamples. Its PXRD pattern is very similar to those of its isomorphs F,G, H, M and N. Like F and G, the powder samples have adehydration/desolvation endotherm at 115-125° C.

Form J is prepared by dissolving azithromycin in 4 volumes of n-propanolat a temperature of about 25-55° C. Water, about 6-7 volumes, is addedat room temperature and the slurry is continuously stirred for 0.5-2hours. The solid form J azithromycin is collected by filtration anddried.

Form K: The PXRD pattern of form K was found in a mixture ofazithromycin form A and microcrystalline wax after annealing at 95° C.for 3 hours. It is a lower hydrate of form A and is a metastable hightemperature form.

Form L: This form has only been observed upon heating the dihydrate;form A. In variable temperature powder X-ray diffraction (VT-PXRD)experiments, a new powder X-ray diffraction pattern appears when form Ais heated to about 90° C. The new form, designated form L, is a lowerhydrate of form A because form A loses about 2.5 weight % at 90° C. byTGA, thus corresponding to a conversion to a monohydrate. When cooled toambient temperature, form L rapidly reverts to form A.

Form M: Isolated from an isopropanol/water slurry, form M incorporatesboth water and isopropanol. Its PXRD pattern and ss-NMR spectrum arevery similar to those of Family I isomorphs, indicating that it belongsto Family I. By analogy to the known crystal structures of Family Iisomorphs, the single crystal structure of form M would be amonohydrate/hemi-isopropranolate. The dehydration/desolvationtemperature of form M is about 115-125° C.

Form M may be prepared by dissolving azithromycin in 2-3 volumes ofisopropanol (IPA) at 40-50° C. The solution is cooled to below 15° C.,preferably below 10° C., more preferably about 5° C. and 2-4 volumes ofcold water about 5° C. are added to effect precipitation. Seeds of formM crystals may be added at the onset of crystallization. The slurry isstirred less than about 5 hours, preferably less than about 3 hours,more preferably less than about 1 hour and most preferably about 30minutes or less and the solids are collected by filtration. The solidsmay be reslurried in isopropanol. This procedure provides form Msubstantially in the absence of azithromycin dihydrate.

Form N: Isolated from water/ethanol/isopropanol slurry of form A, form Ncrystals may contain variable amounts of the crystallization solventsand water. Its water content varies from about 3.4 to about 5.3 weightpercent. Analysis by GC Headspace reveals a variable solvent content ofethanol and isopropanol. The total solvent content of form N samples isusually lower than about 5% depending on the conditions of preparationand drying. The PXRD pattern of form N is similar to that of forms F, G,H, J and M of the Family I isomorphs. The dehydration/desolvationendotherm(s) of the samples of form N may be broader and may varybetween 110-130° C.

Form N azithromycin may be prepared by recrystallizing azithromycin froma mixture of azithromycin crystal latice-incorporating organic solventsand water, such as ethanol, isopropanol, n-propanol, acetone,acetonitirile etc. The solvent mixture is heated to 45-60° C. andazithromycin is added to the heated solvent mixture, up to a total ofabout 4 volumes. Upon dissolution, 1-3 volumes of water are added withcontinuous agitation at 45-60° C. Form N azithromycin precipitates as awhite solid. The slurry is allowed to cool to ambient temperature withstirring. Solid form N azithromycin is isolated by filtration and dried.

Form O: This crystal form is a hemihydrate hemi-n-butanol solvate ofazithromycin free base by single crystal structural data (Table 8A). Itwas isolated from n-butanol solution of azithromycin with diffusion ofantisolvent. The crystal structure of form O is isomorphic to crystalforms of Family I.

Azithromycin is completely dissolved in n-butanol. Addition of anantisolvent, such as hexane, water, IPE or other non-solvent, bydiffusion results in precipitation of Form O.

Form P: This is a proposed crystal form, being a hemihydratehemi-n-pentanol solvate of azithromycin free base. It can be isolatedfrom an n-pentanol solution of azithromycin with diffusion of anantisolvent. The crystal structure of form P is isomorphic to crystalforms of Family I.

Form P of azithromycin may be prepared as following: Azithromycin iscompletely dissolved in n-pentanol; addition of an antisolvent, such ashexane, water, isopropyl ether (IPE) or other non-solvent, by diffusionresults in precipitation of Form P.

Form Q: The crystal form of Q exhibits a unique powder X-ray diffractionpattern. It contains about 4% water and about 4.5% THF, being a hydratehemi THF solvate. The main dehydration/desolvation temperature is fromabout 80 to about 110° C.

Azithromycin dihydrate is dissolved in 6 volumes of THF and 2 volumes ofwater are added. The solution is allowed to evaporate to dryness atambient conditions to afford crystalline Form Q.

Form R: This crystalline form is prepared by adding amorphousazithromycin to 2.5 volumes of tert-butyl methyl ether (MTBE). Theresulting thick white suspension is stirred 3 days at ambientconditions. Solids are collected by vacuum filtration and air dried. Theresulting bulk azithromycin form R has a theoretical water content of2.1 weight % and a theoretical methyl tert-butyl ether content of 10.3weight %.

Due to the similarity in their structures, isomorphs have propensity toform a mixture of the forms within a family, sometimes termed as ‘mixedcrystals’ or ‘crystalline solid solution’. Form N is such a solidcrystalline solution and was found to be a mixture of Family I isomorphsby solvent composition and solid-state NMR data.

Both Family I and Family II isomorphs are hydrates and/or solvates ofazithromycin. The solvent molecules in the cavities have tendency toexchange between solvent and water under specific conditions. Therefore,the solvent/water content of the isomorphs may vary to a certain extent.

The crystal forms of isomorphic Family I are more stable than form Awhen subjected to heating. Forms F, G, H, J, M and N showed higher onsetdehydration temperatures at 110-125° C. than that of form A with anonset dehydration temperature at about 90 to about 110° C. andsimultaneous solid-state conversion to form L at about 90° C.

Amorphous azithromycin: All crystal forms of azithromycin contain wateror solvent(s) or both water and solvent(s). When water and solvent(s)are removed from the crystalline solids, azithromycin becomes amorphous.Amorphous solids have advantages of high initial dissolution rates.

The starting material for the synthesis of the various crystal forms inthe examples below was azithromycin dihydrate unless otherwise noted.Other forms of azithromycin such as amorphous azithromycin or othernon-dihydrate crystalline forms of azithromycin may be used.

EXAMPLES Example 1 Preparation of Form D

Form D was prepared by slurrying azithromycin dihydrate in cyclohexanefor 2-4 days at an elevated temperature, e.g. 25-50° C. The crystallinesolids of form D were collected by filtration and dried.

Example 2 Preparation of Form F

2A: Azithromycin dihydrate was slowly added to one volume of warmethanol, about 70° C., and stirred to complete dissolution at 65 to 70°C. The solution was allowed to cool gradually to 2-5° C. and one volumeof chilled water was added The crystalline solids were collected shortly(preferably less than 30 minutes) after addition of water by vacuumfiltration.

2B: Azithromycin dihydrate is slowly added to one volume of warmethanol, about 70° C., and stirred to complete dissolution at 65 to 70°C. The solution is allowed to cool gradually to 2-5° C. and ethanolvolume may be reduced by vacuum distillation. Seeds of Form F 1-2% wtmay be introduced to facilitate the crystallization. After stirring upto 2 hours the crystalline solids are collected by vacuum filtration.The isolation of the crystals yields substantially pure form Fazithromycin, form F azithromycin substantially free of form Gazithromycin and form F azithromycin substantially free of azithromycindihydrate.

Example 3 Preparation of Form G

A reaction vessel was charged with form A azithromycin. In a separatevessel, 1.5 volumes methanol and 1.5 volumes water were mixed. Thesolvent mixture was added to the reaction vessel containing the form Aazithromycin. The slurry was stirred with heating to 50° C. forapproximately 5 hours. Heating was discontinued and the slurry wasallowed to cool with stirring to ambient temperature. The form Gazithromycin was collected by filtration and allowed to air dry forapproximately 30 minutes. The collected form G azithromycin was furtherdried in a vacuum oven at 45° C. This procedure yields substantiallypure form G azithromycin, and form G azithromycin substantially free ofazithromycin dihydrate.

Example 4 Preparation of Form J

Form J was prepared by dissolving azithromycin in 4 volumes ofn-propanol at a temperature of about 25° C. Water (6.7 volumes) wasadded and the slurry is continuously stirred for 1 hour, followed bycooling to about 0° C. The solid form J azithromycin was collected byfiltration and dried.

Example 5 Preparation of Form M Substantially in the Absence ofAzithromycin Dihydrate

5A: Azithromycin dihydrate is completely dissolved in 2 volumes of warmisopropanol 40-50° C. Seeds of Form M may be optionally introduced tofacilitate the crystallization. The solution is then cooled to 0-5° C.and 4 volumes of chilled water as antisolvent are added and the solidsare collected by vacuum filtration. The solids are reslurried in 1volume of isopropanol for 3-5 hours at 40-45° C. and then cooled to 0-5°C. The crystalline solids are collected shortly (about 15 minutes) afteraddition of water by vacuum filtration. The solids are reslurried in 0.5to 1 volume of isopropanol at 25-40° C. and cooled to about 5° C.followed by filtration to collect solids of form M.

These procedures yield substantially pure form M azithromycin, form Mazithromycin substantially free of form G azithromycin and form Mazithromycin substantially free of azithromycin dihydrate

Example 6 Preparation of Form N

Two volumes of ethanol and 2 volumes of isopropanol were added to areaction vessel and heated to 50° C. Azithromycin form A was added withstirring to the heated ethanol/isopropanol mixture to yield a clearsolution. The reaction vessel was charged with 2 volumes distilled water(ambient temperature). Stirring was continued at 50° C. and solid form Nazithromycin precipitated after approximately 1 hr. Heating wasdiscontinued 5 hours after the addition of the water. The slurry wasallowed to cool to ambient temperature. Precipitated form N azithromycinwas collected by filtration and dried for 4 hours in vacuum oven at 45°C.

Example 7 Preparation of Amorphous Azithromycin

Crystalline form A azithromycin was heated to 110-120° C. in an oven forovernight under vacuum. The amorphous solids were collected and storedwith desiccant as needed.

Example 8 Preparation of Form H

Azithromycin dihydrate or other crystal forms was dissolved in 6 volumesof propylene glycol. To the resulting propylene glycol solution ofazithromycin, 2 volumes of water were added and precipitation occurred.The slurry was stirred for 24 hours and the solids were filtered andair-dried at ambient temperature to afford crystalline Form H.

Example 9 Preparation of Form Q

The crystalline powder was prepared by dissolving 500 mg azithromycinForm A in 2 ml THF. To the clear, colorless solution at room temperaturewas added 1 ml water. When the solution became cloudy an additional 1 mlTHF was added to dissolve the azithromycin completely, and the solutionwas stirred at ambient temperature. Solvent was allowed to evaporateover 7 days, after which the dry solids were collected andcharacterized.

Example 10 Powder X-Ray Diffraction Analysis

Powder patterns were collected using a Bruker D5000 diffractometer(Madison, Wis.) equipped with copper radiation, fixed slits (1.0, 1.0,0.6 mm), and a Kevex solid state detector. Data was collected from 3.0to 40.0 degrees in 2 theta using a step size of 0.04 degrees and a steptime of 1.0 seconds. The results are summarized in Table 9.

The experimental PXRD diffraction pattern of azithromycin form A isgiven in FIG. 2.

The experimental PXRD diffraction pattern of azithromycin form D isgiven in FIG. 6.

The experimental PXRD diffraction pattern of azithromycin form F isgiven in FIG. 10.

The experimental PXRD diffraction pattern of azithromycin form G isgiven in FIG. 13.

The experimental PXRD diffraction pattern of azithromycin form J isgiven in FIG. 16.

The experimental PXRD diffraction pattern of azithromycin form M isgiven in FIG. 18.

The experimental PXRD diffraction pattern of azithromycin form N isgiven in FIG. 19.

The experimental PXRD diffraction pattern of amorphous azithromycin isgiven in FIG. 20.

The experimental PXRD diffraction pattern of azithromycin form Q isgiven in FIG. 30.

The experimental PXRD diffraction pattern of azithromycin form R isgiven in FIG. 31.

The experimental variability from sample to sample is about ±0.2° in 2theta, and the same variations were observed between the calculatedpowder from single crystal structure and experimental data. Detailedanalysis showed that the isomorphs in Family I can be discerned by PXRDwith sets of characteristic peaks given in Table 9. TABLE 9 AzithromycinPowder X-ray Diffraction Peaks in 2-theta ±0.2° A D F G J M N Q  7.2  3.9  5.7  5.0  5.0  5.0   6.2  5.7  7.9  7.3   6.2  5.8  5.7  5.6  7.3 6.1  9.3  7.7  7.4   6.2   6.2   6.2  7.8   6.8  9.9 10.1  7.8  7.4 7.3  7.3  9.8   8.4 11.2 10.6  8.9  7.9  7.8  7.8

 9.5 12.0 11.5  9.8  9.8  8.2  8.2 11.9 10.6 12.7 12.3 10.3 10.2  9.7 9.8 12.5 11.2 13.0 12.8

10.8 10.3 10.2

11.5 14.0 13.6

12.4 15.6 14.5 11.9

11.9

12.7 16.0 15.4 12.2 12.0 11.9 12.2 15.3 13.4 16.4 15.6 12.5 12.5 12.312.5 15.7 13.6 16.8 16.9

13.3 12.5

14.1 17.5 18.3

14.4 18.2 19.0

15.3

14.9 18.7 19.9

16.3 19.1 20.8 15.3

15.3

18.5 17.2 19.8 21.4 15.7 15.3 15.7

19.0 18.2 20.5 21.6

15.7

19.6 19.0 20.9 22.0

18.4 20.0 19.5 21.2 23.0

18.5 20.4 19.8 21.6 23.3

19.1 21.0 20.2 21.8

19.6 21.8 20.5 24.0 18.0

18.1 20.0 22.5 21.1 18.5 18.1 18.5 20.4 23.5 21.6 19.0 18.6 19.0 20.921.9 19.6 19.0 19.7 21.7 22.2 20.0 19.6 20.0 22.5 23.6 20.5 20.0 20.423.2 25.1 21.0 20.5 20.9 23.6 21.7 21.1 21.7 22.0 21.8 22.4 22.4 22.522.6 22.6 23.5 23.3 23.1 23.5 23.5 A D F G J M N QThe peaks underlined are the characteristic peaks among forms A, D,Family I and Q.The peaks in italic and underlined are the sets of peaks that arecharacteristic within Family I isomorphs.

The peaks underlined are the characteristic peaks among forms A, D,Family I and Q.

The peaks in italic and underlined are the sets of peaks that arecharacteristic within Family I isomorphs.

Family I isomorphs have the following common characteristics: thediffraction peaks at 6.2, 11.2, 21.0±0.1 and 22.5±0.1 degree in 2-theta.Each isomorph displays representative sets of diffraction peaks given inthe following, and each set has characteristic spacing between thepeaks.

The diffraction peak positions reported are accurate to within ±0.2degree of 2-theta.

A representative PXRD pattern of form A is shown in FIG. 2. Form Adisplays peaks at 9.3, 13.0 and 18.7 degrees of 2-theta.

A representative PXRD pattern of form D is shown in FIG. 6. Form Ddisplays peaks at 3.9, 10.1, 10.6 and 21.4 degrees of 2-theta.

A representative PXRD pattern of Form F is shown in FIG. 10. Form Fdisplays the characteristic peaks of Family I and three sets of peaks,being set 1 at 2-theta of 11.2 and 11.5; set 2 at 2-theta of 13.9, 14.3,14.7 and 14.8; set 3 at 2-theta of 16.2, 16.6, 17.1, 17.2 and 17.7.

A representative PXRD pattern of Form G is shown in FIG. 13. Form Gdisplays the characteristic peaks of Family I and three sets of peaks,being set 1 at 2-theta of 11.2 and 11.6 2; set at 2-theta of 14.0, 14.4,14.6 and 14.9; set 3 at 2-theta of 16.3, 16.6, 17.2, 17.4 and 17.8.

A representative PXRD pattern of Form J is shown in FIG. 16. Form Jdisplays the characteristic peaks of Family I and three sets of peaks,being set 1 at 2-theta of 11.2 and 11.4; set 2 at 2-theta of 13.9, 14.2and 14.6; set 3 at 2-theta of 16.0, 16.6, 17.0, 17.2 and 17.5.

A representative PXRD pattern of Form M is shown in FIG. 18. Form Mdisplays the characteristic peaks of Family I and three sets of peaks,being set 1 at 2-theta of 11.2; set 2 at 2-theta of 14.0 and 14.6; set 3at 2-theta of 15.9, 16.6, 17.1 and 17.5.

A representative PXRD pattern of Form N is shown in FIG. 10. Form Ndisplays the characteristic peaks of Family I. The sets of peaks of formN are similar to those of forms F, G, J and M, being set 1 at 2-theta of11.2 to 11.6; set 2 at 2-theta of 13.9 to 15.0; and set 3 at 2-theta of15.9 to 17.9, with the peaks may vary slightly in position, intensityand width due to mixing of variable proportion of isomorphs in Family I.

A representative PXRD pattern of form Q is shown in FIG. 30. Form 0displays peaks at 2-theta of 6.8, 8.4 and 20.2 degree.

A representative PXRD pattern of form R is shown in FIG. 31.

Example 11 Single Crystal X-Ray Analysis

Data were collected at room temperature using Bruker X-raydiffractometers equipped with copper radiation and graphitemonochromators. Structures were solved using direct methods. The SHELXTLcomputer library provided by Bruker AXS, Inc facilitated all necessarycrystallographic computations and molecular displays (SHELXTL™ ReferenceManual, Version 5.1, Bruker AXS, Madison, Wis., USA (1997)).

Example 12 Calculation of PXRD Pattern from Single Crystal Data

To compare the results between a single crystal and a powder sample, acalculated powder pattern can be obtained from single crystal results.The XFOG and XPOW computer programs provided as part of the SHELXTLcomputer library were used to perform this calculation. Comparing thecalculated powder pattern with the experimental powder pattern confirmswhether a powder sample corresponds to an assigned single crystalstructure (Table 9A). This procedure was performed on the crystal formsof azithromycin A, D, F, G, and J.

The calculated PXRD diffraction pattern of azithromycin form A is givenin FIG. 1.

The calculated PXRD diffraction pattern of azithromycin form D is givenin FIG. 5.

The calculated PXRD diffraction pattern of azithromycin form F is givenin FIG. 9.

The calculated PXRD diffraction pattern of azithromycin form G is givenin FIG. 12.

The calculated PXRD diffraction pattern of azithromycin form J is givenin FIG. 15.

The results are displayed in the overlaid powder X-ray diffractionpatterns for forms A, D, F, G, and J in FIGS. 3, 7, 11, 14 and 17,respectively. The lower pattern corresponds to the calculated powderpattern (from single crystal results) and the upper pattern correspondsto a representative experimental powder pattern. A match between the twopatterns indicated the agreement between powder sample and thecorresponding single crystal structure. TABLE 9A Cacluated andExperimental PXRD Peaks of Isomorphs of Family I F G J M F experi- Gexperi- J experi- experi- calculated mental calculated mental calculatedmental tal 5.2 5.0 5.7 5.8 5.8 5.7 5.6 6.3 6.2 6.2 6.2 6.3 6.2 6.2 7.47.4 7.5 7.4 7.4 7.3 7.3 7.9 7.8 7.9 7.9 7.9 7.8 7.8 8.8 8.9 8.9 9.3 8.38.2 8.2 9.9 9.8 9.9 9.9 9.8 9.7 9.8 10.3 10.3 10.2 10.4 10.3 10.2 10.910.9 10.8 11.3 11.2 11.3 11.2 11.2 11.2 11.2 11.5 11.4 11.6 11.6 11.411.4 missing 12.0 11.9 12.0 11.9 12.0 11.9 11.9 12.3 12.2 12.3 12.3 12.312.2 12.6 12.5 12.5 12.5 12.6 12.5 12.5 14.0 14.0 13.4 13.3 14.0 13.914.0 14.3 14.3 14.1 14.0 14.2 14.2 missing 14.4 14.4 14.7 14.7 14.7 14.614.7 14.6 14.6 14.9 14.8 14.9 14.9 14.8 15.4 15.3 15.4 15.3 15.3 15.315.3 15.8 15.7 15.7 15.7 15.8 15.7 15.9 16.2 16.2 16.3 16.3 16.0 16.0missing 16.6 16.6 16.6 16.6 16.7 16.6 16.6 17.1 17.2 17.1 17.1 17.0 17.117.3 17.3 17.3 17.2 17.4 17.2 missing 17.5 17.4 17.5 17.4 17.6 17.5 17.517.7 17.7 17.9 17.8 17.9 18.0 18.0 18.1 18.1 18.2 18.1 18.4 18.6 18.518.7 18.7 18.5 18.5 18.5 19.1 19.0 19.1 19.0 19.1 19.0 19.1 19.7 19.619.6 19.6 19.8 19.7 19.6 20.0 20.0 20.0 20.0 20.1 20.0 20.0 20.5 20.420.6 20.5 20.5 20.4 20.4 21.1 21.0 21.2 21.0 20.8 20.9 20.9 21.8 21.721.6 21.6 21.7 21.7 22.1 22.0 21.8 21.8 21.8 22.5 22.4 22.3 22.2 22.522.4 22.5 22.7 22.6 22.5 22.5 22.8 22.6 23.1 23.1 22.9 23.4 23.3 23.223.6 23.5 23.5 23.5 23.7 23.5 23.6

Example 13 Solid State NMR Analysis

Solid State NMR Analysis:

All ¹³C solid state NMR spectra were collected on an 11.75 Tspectrometer (Bruker Biospin, Inc., Billerica, Mass.), corresponding to125 MHz ¹³C frequency. The spectra were collected using across-polarization magic angle spinning (CPMAS) probe operating atambient temperature and pressure. Depending on the quantity of sampleanalyzed, 7 mm BL or 4 mm BL Bruker probes were employed, accomodating300 mg and 75 mg of sample with maximum speeds of 7 kHz and 15 kHz,respectively. Data were processed with an exponential line broadeningfunction of 5.0 Hz. Proton decoupling of 65 kHz and 100 kHz were usedwith the 7 mm and 4 mm probes, respectively. A sufficient number ofacquisitions were averaged out to obtain adequate signal-to-noise ratiosfor all peaks. Typically, 600 scans were acquired with recycle delay of3.0 s (seconds), corresponding approximately to a 30 minute totalacquisition time. Magic angle was adjusted using KBr powder according tostandard NMR vendor practices. The spectra were referenced relative toeither the methyl resonace of hexamethylbenzen (HMB) at 17.3 ppm or theupfield resonance of adamantane (ADM) at 29.5 ppm. HMB referencedspectra show chemical shifts of all peaks shifted down field by 0.08 ppmwith respect to same spectra referenced to ADM. The spectral windowminimally included the spectra region from 190 to 0 ppm. The results aresummarized in Table 10. Ss-NMR spectra for forms M, H and R werereferenced to ADM. Ss-NMR spectra for forms A, D, G, F, J and N werereferenced to HMB. Forms H and R were spun at a rate of 15 kHz. TABLE 10¹³C ss-NMR chemical shifts of Azithromycin (±0.2 ppm) A D G F J M N H R178.1  178.1  179.5* 179.5  179.6  179.6  179.6  179.5  177.9  104.1 103.9  105.5  178.6  178.4  105.6  178.7  178.7  104.6  98.4 95.1 103.5 105.5  105.5  103.4  105.6  105.4  103.6  84.6 84.2 95.0 103.4  103.4 94.9 103.6  103.2  95.3 82.6 79.4 86.2 94.9 95.0 86.7 95.0 95.0 85.479.3 78.9 83.1 86.4 86.4 82.9 86.5 86.4 84.0 78.3 75.7 78.9 83.0 82.979.3 83.1 82.1 79.4 75.6 74.6 78.2 79.1 79.2 78.1 79.0 79.2 79.0 74.774.0 77.6 78.1 78.1 77.0 77.9 78.3 75.6 73.9 72.9 76.4 77.9 76.8 76.776.5 78.0 74.5 73.5 71.9 75.7 76.5 76.2 74.7 74.8 76.4 73.9 70.8 71.074.7 74.7 74.1 74.2 74.2 74.7 73.9 68.0 69.4 74.3 74.1  74.1** 71.3 73.674.1 72.9 66.2 67.8 73.5 73.5 72.0 69.2 71.5 73.5 71.8 63.8 65.7 71.371.4 71.3 68.6 69.2 73.1 71.0 63.2 64.7 69.1 69.1 69.2 67.3 68.7 71.269.1 52.2 49.2 68.8 68.6 68.6 66.2 67.3 69.1 67.5 44.3 45.8 67.4 67.3 67.3** 65.5 66.2 68.4 65.6 42.6 43.1 65.9 66.1  66.2** 63.8 65.7 67.364.5 41.7 40.6 65.2 65.6  65.5** 63.3 63.7 66.9 49.4 39.1 37.1 64.0 63.663.7 50.0 58.1 66.1 45.7 35.4 36.4 63.3 58.0 50.0 47.1 50.1  65.5* 42.934.6 29.6 50.0 50.0 46.9 45.9 47.1  63.7* 41.6 26.9 29.3 46.9 47.0 45.944.7 46.0 49.9 40.4 26.3 28.0 46.0 45.9 44.7 43.8 44.8 46.8 37.0 23.727.7 44.5 44.7 43.7 41.9 43.8 45.9 36.2 23.3 22.1 43.7 43.7 41.6 41.141.5 44.5 29.4 21.7 21.1 41.5 41.5 41.0 37.4 41.1  43.8* 29.0 19.5 18.640.8 41.1 37.1 36.2 37.3 41.7 28.2 17.5 16.7 37.5 37.3  36.5** 33.6 36.540.9 27.4 15.9 16.1 36.5 36.4  35.4** 30.1 33.7 37.1 21.4 13.2 10.6 33.633.6 33.5 28.1 30.4 36.3 20.8 11.3  9.0 30.0 30.3 30.4 27.2 28.1 33.718.7  7.2  8.6 27.9 28.0 28.0 26.0 27.2 33.3 16.5 27.3 27.1 27.1 23.226.0  30.5* 16.1 23.1 23.2 25.2 22.8 23.2 27.9 15.7 22.5 22.6 23.2 22.522.6 27.1 10.3 21.9 21.9  22.5** 21.8 22.0 23.1  9.6 20.9 20.8  21.9**20.2 20.8 22.6  8.9 20.2 20.4 20.7 18.9 19.0 22.3  8.6 18.8 18.9 18.917.4 16.9 21.9 17.0 16.8 16.8 16.3 15.8 20.7 16.0 17.2  15.6** 15.5 12.220.3 12.2 15.7 12.1 12.1  9.9 18.8 10.4 12.2 11.5 10.3  9.4 17.1  9.910.1 12.1  9.6  7.9 16.6  9.3  9.8 10.0  9.3  6.6 15.8  7.6  9.3  9.3 7.7 15.4  6.5  7.9  8.1  7.1 12.0  6.6   6.8**  9.9  9.1  7.9  7.0The chemical shifts labeled in bold and underlined are the peaks or setsof peaks representative of each form.The chemical shifts labeled in italic are the solvent peaks that may bebroad and variable (±0.4 ppm).The chemical shifts labeled with single asterisk may show splitting of<0.3 ppm.The chemical shifts labeled with double asterisk may show variation of±0.3 ppm

The chemical shifts reported are accurate to within ±0.2 ppm unlessotherwise indicated.

A representative ¹³C ssNMR spectrum of form A is shown in FIG. 21. FormA displays a peak at 178.1 ppm, and peaks at 104.1, 98.4, 84.6, 26.9,13.2, 11.3 and 7.2 ppm.

A representative ¹³C ssNMR spectrum of form D is shown in FIG. 22. FormD displays the highest chemical shift peak of 178.1 ppm and peaks atchemical shifts of 103.9, 95.1, 84.2, 10.6, 9.0 and 8.6 ppm.

A representative ¹³C ssNMR spectrum of form F is shown in FIG. 23. FormF has two chemical shift peaks at approximately 179.1±2 ppm, being 179.5ppm and 178.6 ppm, and a set of 5 peaks at 10.1, 9.8, 9.3, 7.9, and 6.6ppm, and ethanol peaks at 58.0±0.5 ppm and 17.2±0.5 ppm. The solventpeaks can be broad and relatively weak in intensity.

A representative ¹³C ssNMR spectrum of form G is shown in FIG. 24. FormG has the highest chemical shift peak of 179.5 ppm, being a single peakwith possible splitting of <0.3 ppm and a set of 5 peaks at 10.4, 9.9,9.3, 7.6, 6.5 ppm.

A representative ¹³C ssNMR spectrum of form J is shown in FIG. 25. FormJ has two chemical shift peaks at approximately 179.1±2 ppm, those being179.6 ppm and 178.4 ppm, a set of 4 peaks at 10.0, 9.3, 8.1 and 6.8 ppmand n-propanol peaks at 11.5±0.5 ppm and 25.2±0.5 ppm. The solvent peakcan be broad and relatively weak in intensity.

A representative ¹³C ssNMR spectrum of form M is shown in FIG. 26. FormM has one chemical shift peak at 179±1 ppm, being 179.6 ppm, peaks at41.9, and 16.3 ppm, a set of 5 peaks at 10.3, 9.6, 9.3, 7.7 and 7.1 ppmand an isopropanol peak at 26.0±0.5 ppm. The solvent peak can be broadand relatively weak in intensity.

A representative ¹³C ssNMR spectrum of form N is shown in FIG. 27. FormN displays chemical shifts as a combination of isomorphs in Family I.The peaks may vary in chemical shift and in relative intensities andwidth due to the mixing of variable proportion of isomorphs contained inthe form N crystalline solid solution.

A representative ¹³C ssNMR spectrum of amorphous form is shown in FIG.28. The amorphous azithromycin displays broad chemical shifts. Thecharacteristic chemical shifts have the peak positions at 179 and 11±0.5ppm.

A summary of the observed ssNMR peaks for forms A, D, F, G, H, J, M, Nand R azithromycin is given in Table 10.

Example 14 NMR Analysis of a Dosage Form

To demonstrate the ability of ¹³C ssNMR to identify the form ofazithromycin contained in a pharmaceutical dosage form, coatedazithromycin tablets containing form G azithromycin were prepared andanalyzed by ¹³C ssNMR. Tablets were wet granulated and tabletted on anF-Press (Manesty, Liverpool, UK) using 0.262″×0.531″ tooling. Tabletswere formulated and tabletted to contain 250 mg of form G azithromycinwith a total tablet weight of 450 mg using the formula given below. Thetablets were uniformly coated with pink Opadry II® (mixture of lactosemonohydrate, hydroxypropylmethylcellulose, titanium dioxide, Drug &Cosmetic red # 30, and triacetin) (Colorcon, West Point, Pa.). MaterialPercentage Batch(g) Azithromycin form “G” 58.23 174.69 Pregellatinizedcorn starch 6.00 18.00 Anhydrous dicalcium phosphate 30.85 92.55 Sodiumcroscarmelose 2.00 6.00 Magnesium stearate with 10% sodium 2.92 8.76laurel sulfate Total 100.00 300.00

A coated tablet was gently crushed and the powdered sample was packedwith a packing tool in solid state rotor containing no ¹³C background.Analysis of the sample was performed under conditions outlined inExample 13.

A representative ¹³C ssNMR spectrum of the tablet containing form Gazithromycin is given in FIG. 29.

Example 15 Antimicrobial Activity

The activity of the crystal forms of the present invention againstbacterial and protozoa pathogens is demonstrated by the compound'sability to inhibit growth of defined strains of human (Assay I) oranimal (Assays II and III) pathogens.

Assay I

Assay I, described below, employs conventional methodology andinterpretation criteria and is designed to provide direction forchemical modifications that may lead to compounds that circumventdefined mechanisms of macrolide resistance. In Assay I, a panel ofbacterial strains is assembled to include a variety of target pathogenicspecies, including representatives of macrolide resistance mechanismsthat have been characterized. Use of this panel enables the chemicalstructure/activity relationship to be determined with respect topotency, spectrum of activity, and structural elements or modificationsthat may be necessary to obviate resistance mechanisms. Bacterialpathogens that comprise the screening panel are shown in the tablebelow. In many cases, both the macrolide-susceptible parent strain andthe macrolide-resistant strain derived from it are available to providea more accurate assessment of the compound's ability to circumvent theresistance mechanism. Strains that contain the gene with the designationof ermA/ermB/ermC are resistant to macrolides, lincosamides, andstreptogramin B antibiotics due to modifications (methylation) of 23SrRNA molecules by an Erm methylase, thereby generally prevent thebinding of all three structural classes. Two types of macrolide effluxhave been described; msrA encodes a component of an efflux system instaphylococci that prevents the entry of macrolides and streptograminswhile mefA/E encodes a transmembrane protein that appears to efflux onlymacrolides. Inactivation of macrolide antibiotics can occur and can bemediated by either a phosphorylation of the 2′-hydroxyl (mph) or bycleavage of the macrocyclic lactone (esterase). The strains may becharacterized using conventional polymerase chain reaction (PCR)technology and/or by sequencing the resistance determinant. The use ofPCR technology in this application is described in J. Sutcliffe et al.,“Detection Of Erythromycin-Resistant Determinants By PCR”, AntimicrobialAgents and Chemotherapy, 40(11), 2562-2566 (1996). The assay isperformed in microtiter trays and interpreted according to PerformanceStandards for Antimicrobial Disk Susceptibility Tests—Sixth Edition:Approved Standard, published by The National Committee for ClinicalLaboratory Standards (NCCLS) guidelines; the minimum inhibitoryconcentration (MIC) is used to compare strains. The crystalline compoundis initially dissolved in dimethylsulfoxide (DMSO) as 40 mg/ml stocksolution. Strain Designation Macrolide Resistance Mechanism(s)Staphylococcus aureus 1116 susceptible parent Staphylococcus aureus 1117ErmB Staphylococcus aureus 0052 susceptible parent Staphylococcus aureus1120 ErmC Staphylococcus aureus 1032 msrA, mph, esterase Staphylococcushemolyticus 1006 msrA, mph Streptococcus pyogenes 0203 susceptibleparent Streptococcus pyogenes 1079 ErmB Streptococcus pyogenes 1062susceptible parent Streptococcus pyogenes 1061 ErmB Streptococcuspyogenes 1064 ErmB Streptococcus agalactiae 1024 susceptible parentStreptococcus agalactiae 1023 ErmB Streptococcus pneumoniae 1016Susceptible Streptococcus pneumoniae 1046 ErmB Streptococcus pneumoniae1095 ErmB Streptococcus pneumoniae 1175 MefE Streptococcus pneumoniae0085 Susceptible Haemophilus influenzae 0131 Susceptible Moraxellacatarrhalis 0040 Susceptible Moraxella catarrhalis 1055 erythromycinintermediate resistance Escherichia coli 0266 Susceptible

Assay II is utilized to test for activity against Pasteurella multocidaand Assay III is utilized to test for activity against Pasteurellahaemolytica.

Assay II

This assay is based on the liquid dilution method in microliter format.A single colony of P. multocida (strain 59A067) is inoculated into 5 mlof brain heart infusion (BHI) broth. The test compound is prepared bysolubilizing 1 mg of the compound in 125 μl of dimethylsulfoxide (DMSO).Dilutions of the test compound are prepared using uninoculated BHIbroth. The concentrations of the test compound used range from 200 μg/mlto 0.098 μg/ml by two-fold serial dilutions. The P. multocida inoculatedBHI is diluted with uninoculated BHI broth to make a 10⁴ cell suspensionper 200 μl. The BHI cell suspensions are mixed with respective serialdilutions of the test compound, and incubated at 37° C. for 18 hours.The minimum inhibitory concentration (MIC) is equal to the concentrationof the compound exhibiting 100% inhibition of growth of P. multocida asdetermined by comparison with an uninoculated control.

Assay III

This assay is based on the agar dilution method using a SteersReplicator. Two to five colonies isolated from an agar plate areinoculated into BHI broth and incubated overnight at 37° C. with shaking(200 rpm). The next morning, 300 μl of the fully grown P. haemolyticapreculture is inoculated into 3 ml of fresh BHI broth and is incubatedat 37° C. with shaking (200 rpm). The appropriate amounts of the testcompounds are dissolved in ethanol and a series of two-fold serialdilutions are prepared. Two ml of the respective serial dilution ismixed with 18 ml of molten BHI agar and solidified. When the inoculatedP. haemolytica culture reaches 0.5 McFarland standard density, about 5μl of the P. haemolytica culture is inoculated onto BHI agar platescontaining the various concentrations of the test compound using aSteers Replicator and incubated for 18 hours at 37° C. Initialconcentrations of the test compound range from 100-200 μg/ml. The MIC isequal to the concentration of the test compound exhibiting 100%inhibition of growth of P. haemolytica as determined by comparison withan uninoculated control.

The in vivo activity of the crystal forms of the present invention canbe determined by conventional animal protection studies well known tothose skilled in the art, usually carried out in mice.

Mice are allotted to cages (10 per cage) upon their arrival, and allowedto acclimate for a minimum of 48 hours before being used. Animals areinoculated with 0.5 ml of a 3×10³ CFU/ml bacterial suspension (P.multocida strain 59A006) intraperitoneally. Each experiment has at least3 non-medicated control groups including one infected with 0.1×challenge dose and two infected with 1× challenge dose; a 10× challengedata group may also be used. Generally, all mice in a given study can bechallenged within 30-90 minutes, especially if a repeating syringe (suchas a Cornwall® syringe) is used to administer the challenge. Thirtyminutes after challenging has begun, the first compound treatment isgiven. It may be necessary for a second person to begin compound dosingif all of the animals have not been challenged at the end of 30 minutes.The routes of administration are subcutaneous or oral doses.Subcutaneous doses are administered into the loose skin in the back ofthe neck whereas oral doses are given by means of a feeding needle. Inboth cases, a volume of 0.2 ml is used per mouse. Compounds areadministered 30 minutes, 4 hours, and 24 hours after challenge. Acontrol compound of known efficacy administered by the same route isincluded in each test. Animals are observed daily, and the number ofsurvivors in each group is recorded. The P. multocida model monitoringcontinues for 96 hours (four days) post challenge.

The PD₅₀ is a calculated dose at which the compound tested protects 50%of a group of mice from mortality due to the bacterial infection thatwould be lethal in the absence of drug treatment.

The crystal forms of the present invention (hereinafter “the activecompound(s)”), may be administered through oral, parenteral, topical, orrectal routes in the treatment or prevention of bacterial or protozoainfections. In general, the active compound is most desirablyadministered in dosages ranging from about 0.2 mg per kg body weight perday (mg/kg/day) to about 200 mg/kg/day in single or divided doses (i.e.,from 1 to 4 doses per day), although variations will necessarily occurdepending upon the species, weight and condition of the subject beingtreated and the particular route of administration chosen. However, adosage level that is in the range of about 2 mg/kg/day to about 50mg/kg/day is most desirably employed. Variations may nevertheless occurdepending upon the species of mammal, fish or bird being treated and itsindividual response to said medicament, as well as on the type ofpharmaceutical formulation chosen and the time period and interval atwhich such administration is carried out. In some instances, dosagelevels below the lower limit of the aforesaid range may be more thanadequate, while in other cases still larger doses may be employedwithout causing any harmful side effects, provided that such largerdoses are first divided into several small doses for administrationthroughout the day.

The active compound may be administered alone or in combination withpharmaceutically acceptable carriers or diluents by the routespreviously indicated, and such administration may be carried out insingle or multiple doses. More particularly, the active compound may beadministered in a wide variety of different dosage forms, i.e., they maybe combined with various pharmaceutically acceptable inert carriers inthe form of tablets, capsules, lozenges, troches, hard candies, powders,sprays, creams, salves, suppositories, jellies, gels, pastes, lotions,ointments, sachets, powders for oral suspension, aqueous suspensions,injectable solutions, elixirs, syrups, and the like. Such carriersinclude solid diluents or fillers, sterile aqueous media and variousnon-toxic organic solvents, etc. Moreover, oral pharmaceuticalcompositions can be suitably sweetened and/or flavored. In general, theactive compound is present in such dosage forms at concentration levelsranging from about 1.0% to about 70% by weight.

For oral administration, tablets containing various excipients such asmicrocrystalline cellulose, sodium citrate, calcium carbonate, dicalciumphosphate and glycine may be employed along with various disintegrantssuch as starch (and preferably corn, potato or tapioca starch), alginicacid and certain complex silicates, together with granulation binderslike polyvinylpyrrolidone, sucrose, gelatin and acacia. Additionally,lubricating agents such as magnesium stearate, sodium lauryl sulfate andtalc are often very useful for tabletting purposes. Solid compositionsof a similar type may also be employed as fillers in gelatin capsules;preferred materials in this connection also include lactose or milksugar as well as high molecular weight polyethylene glycols. Whenaqueous suspensions and/or elixirs are desired for oral administration,the active compound may be combined with various sweetening or flavoringagents, coloring matter or dyes, and, if so desired, emulsifying and/orsuspending agents as well, together with such diluents as water,ethanol, propylene glycol, glycerin and various like combinationsthereof.

For parenteral administration, solutions of the active compound ineither sesame or peanut oil or in aqueous propylene glycol may beemployed. The aqueous solutions should be suitably buffered (preferablypH greater than 8) if necessary and the liquid diluent first renderedisotonic. These aqueous solutions are suitable for intravenous injectionpurposes. The oily solutions are suitable for intraarticular,intramuscular and subcutaneous injection purposes. The preparation ofall these solutions under sterile conditions is readily accomplished bystandard pharmaceutical techniques will known to those skilled in theart.

Additionally, it is also possible to administer the active compoundtopically and this may be done by way of creams, jellies, gels, pastes,patches, ointments and the like, in accordance with standardpharmaceutical practice.

For administration to animals other than humans, such as cattle ordomestic animals, the active compounds may be administered in the feedof the animals or orally as a drench composition.

The active compound may also be administered in the form of liposomedelivery systems, such as small unilamellar vesicles, large unilamellarvesicles and multilamellar vesicles. Liposomes can be formed from avariety of phospholipids, such as cholesterol, stearylamine orphosphatidylcholines.

1-138. (canceled)
 139. A crystalline form of azithromycin, wherein saidform is crystalline monohydrate hemi-isopropanol solvate.
 140. Thecrystalline form of azithromycin of claim 139, wherein said formcomprises two water molecules and one isopropanol molecule for each twomolecules of azithromycin.
 141. The crystalline form of azithromycin ofclaim 139, wherein said form is characterized as containing 2-5% ofwater and 1-4% of isopropanol by weight in powder sample.
 142. Thecrystalline form of azithromycin of claim 139, wherein said form ischaracterized as having a ¹³C solid state NMR spectrum comprising atleast one peak with chemical shift of approximately 179±1 ppm.
 143. Thecrystalline form of azithromycin of claim 142, wherein said form ischaracterized as having a ¹³C solid state NMR spectrum comprising atleast one additional peak with chemical shift of approximately 179.6±0.2ppm.
 144. The crystalline form of azithromycin of claim 143, whereinsaid form is characterized as having a ¹³C solid state NMR spectrumcomprising at least one additional peak with chemical shift ofapproximately 41.9±0.2 ppm.
 145. The crystalline form of azithromycin ofclaim 144, wherein said form is characterized as having a ¹³C solidstate NMR spectrum comprising at least one set of additional six peakswith chemical shift between 6.9 to 16.4 ppm.
 146. The crystalline formof azithromycin of claim 139, wherein said form is substantially pure.147. The crystalline form of azithromycin of claim 139, wherein saidform is substantially free of form G azithromycin.
 148. The crystallineform of azithromycin of claim 139, wherein said form is substantiallyfree of azithromycin dihydrate.
 149. A pharmaceutical dosage formcomprising the crystalline form of azithromycin of claim 139 andpharmaceutically acceptable carrier or diluent.
 150. The pharmaceuticaldosage form of claim 149, wherein said dosage form is a tablet,capsules, lozenge, troche, hard candy, powder, spray, cream, salve,suppository, jelly, gel, paste, lotion, ointment, sachet, powder fororal suspension, aqueous suspension, injectable solution, elixir orsyrup.
 151. The pharmaceutical dosage form of claim 150, wherein saiddosage form comprises from about 1.0% to about 70% by weight ofcrystalline monohydrate hemi-isopropanol solvate.
 152. A method oftreating a bacterial infection or a protozoa infection in a mammal,fish, or bird which comprises administering to said mammal, fish or birda therapeutically effective amount of crystalline form of azithromycinaccording to claim 139.